Showing papers in "Analytical Chemistry in 2023"

The New Era of High-Throughput Nanoelectrochemistry

Abstract: ACCESS Metrics & More Article Recommendations ■ CONTENTS High-Throughput Nanopores 320 Nanopore Working Principle 320 Arrayed Nanopore Configurations 321 Machine-Learning Assisted Nanopore Readout 321 Integration Modalities for Nanopores 321 Nanopore-Confined Electrochemistry 322 High-Throughput Scanning Ion Conductance Microscopy 324 From Nanopore to Nanoprobe 324 High-Throughput SICM 324 High-Throughput Imaging 325 High-Throughput Single-Cell Manipulation and Measurements 326 SICM as an Electrochemical Probe 326 Applications in Material Sciences 327 Applications in Life Sciences 328 High-Throughput Scanning Electrochemical Cell Microscopy 329 Technical and Theoretical Developments 329 Electrochemical Measurements and Characterization 331 Popular Redox Reactions and Electrode Materials 331 Corrosion 332 Phase Formation 332 Two-Dimensional Materials 333 Photoelectrochemistry 335 Electrocatalysis: Single Particles and PseudoSingle-Crystal Screening of Structure−Activity 336 Battery Electrode Materials 339 Optical Microscopies in Electrochemistry 339 Overview of Operational Principles 339 Operational Principles 339 Methodologies for Quantitative Image Analysis 340 Converting Local Optical Information into an Electrochemically Relevant Signal 340 Computing and Automatized Image Analysis 340 Imaging Single Events 340 A Complement to Single Nanoobject Electrochemistry 340 Electron Transfer 341 Probing Concentration Profiles 341 Conversion 341 Growth and Dissolution 342 Catalysis and Motion 342 Competing Processes 343 Electrochemistry versus Physical Transformation 343 Competing Electrochemical Reactions 344 One versus Many 344 Seeing Collective Behaviors 344 How to Access Missing Pieces of Information 344 Hyphenation with Local Complementary Information 345 Other Electrochemical and Electronic HighThroughput Imaging Techniques 345 Conclusion 346 Author Information 347 Corresponding Authors 347 Authors 347 Author Contributions 347 Notes 347 Biographies 347 Acknowledgments 348 List of Abbreviations Used 348 References 349

Microelectromechanical Microsystems-Supported Photothermal Immunoassay for Point-of-Care Testing of Aflatoxin B1 in Foodstuff.

Abstract: Accurate identification of acutely toxic and low-fatality mycotoxins on a large scale in a quick and cheap manner is critical for reducing population mortality. Herein, a portable photothermal immunosensing platform supported by a microelectromechanical microsystem (MEMS) without enzyme involvement was reported for point-of-care testing of mycotoxins (in the case of aflatoxin B1, AFB1) in food based on the precise satellite structure of Au nanoparticles. The synthesized Au nanoparticles with a well-defined, graded satellite structure exhibited a significantly enhanced photothermal response and were coupled by AFB1 antibodies to form signal conversion probes by physisorption for further target-promoted competitive responses in microplates. In addition, a coin-sized miniature NIR camera device was constructed for temperature acquisition during target testing based on advanced MEMS fabrication technology to address the limitation of expensive signal acquisition components of current photothermal sensors. The proposed MEMS readout-based microphotothermal test method provides excellent AFB1 response in the range of 0.5-500 ng g-1 with detection limits as low as 0.27 ng g-1. In addition, the main reasons for the efficient photothermal transduction efficiency of Au with different graded structures were analyzed by finite element simulations, providing theoretical guidance for the development of new Au-based photothermal agents. In conclusion, the proposed portable micro-photothermal test system offers great potential for point-of-care diagnostics for residents, which will continue to facilitate immediate food safety identification in resource-limited regions.

Innovative Vibrational Spectroscopy Research for Forensic Application.

Abstract: ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTInnovative Vibrational Spectroscopy Research for Forensic ApplicationAlexis WeberAlexis WeberDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesSupreMEtric LLC, 7 University Pl. B210, Rensselaer, New York 12144, United StatesMore by Alexis WeberView Biography, Bailey HoplightBailey HoplightDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesMore by Bailey HoplightView Biography, Rhilynn OgilvieRhilynn OgilvieDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesMore by Rhilynn OgilvieView Biography, Claire MuroClaire MuroNew York State Police Forensic Investigation Center, Building #30, Campus Access Rd., Albany, New York 12203, United StatesMore by Claire MuroView Biography, Shelby R. KhandasammyShelby R. KhandasammyDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesMore by Shelby R. KhandasammyView Biography, Luis Pérez-AlmodóvarLuis Pérez-AlmodóvarDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesMore by Luis Pérez-AlmodóvarView Biography, Samuel SearsSamuel SearsDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesMore by Samuel SearsView Biography, and Igor K. Lednev*Igor K. LednevDepartment of Chemistry, University at Albany, SUNY, 1400 Washington Avenue, Albany, New York 12222, United StatesSupreMEtric LLC, 7 University Pl. B210, Rensselaer, New York 12144, United States*E-mail: [email protected]More by Igor K. LednevView Biographyhttps://orcid.org/0000-0002-6504-531XCite this: Anal. Chem. 2023, 95, 1, 167–205Publication Date (Web):January 10, 2023Publication History Published online10 January 2023Published inissue 10 January 2023https://doi.org/10.1021/acs.analchem.2c05094Copyright © 2023 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views485Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (7 MB) Get e-AlertsSUBJECTS:Anatomy,Fibers,Infrared spectroscopy,Mathematical methods,Raman spectroscopy Get e-Alerts

Latest Improvements and Expanding Applications of Solid-Phase Microextraction.

Abstract: ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTLatest Improvements and Expanding Applications of Solid-Phase MicroextractionJuan ZhengJuan ZhengMinistry of Education (MOE) Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry/School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510006, ChinaMore by Juan ZhengView Biography, Yixin KuangYixin KuangMinistry of Education (MOE) Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry/School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510006, ChinaMore by Yixin KuangView Biography, Suxin ZhouSuxin ZhouMinistry of Education (MOE) Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry/School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510006, ChinaMore by Suxin ZhouView Biography, Xinying GongXinying GongMinistry of Education (MOE) Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry/School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510006, ChinaMore by Xinying GongView Biography, and Gangfeng Ouyang*Gangfeng OuyangMinistry of Education (MOE) Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry/School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510006, China*[email protected]More by Gangfeng OuyangView Biographyhttps://orcid.org/0000-0002-0797-6036Cite this: Anal. Chem. 2023, 95, 1, 218–237Publication Date (Web):January 10, 2023Publication History Received26 July 2022Published online10 January 2023Published inissue 10 January 2023https://doi.org/10.1021/acs.analchem.2c03246Copyright © 2023 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views321Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (9 MB) Get e-AlertsSUBJECTS:Adsorption,Coating materials,Extraction,Fibers,Food Get e-Alerts

Robust and Easy-to-Use One-Pot Workflow for Label-Free Single-Cell Proteomics

Abstract: The analysis of ultralow input samples or even individual cells is essential to answering a multitude of biomedical questions, but current proteomic workflows are limited in their sensitivity and reproducibility. Here, we report a comprehensive workflow that includes improved strategies for all steps, from cell lysis to data analysis. Thanks to convenient-to-handle 1 μL sample volume and standardized 384-well plates, the workflow is easy for even novice users to implement. At the same time, it can be performed semi-automatized using CellenONE, which allows for the highest reproducibility. To achieve high throughput, ultrashort gradient lengths down to 5 min were tested using advanced μ-pillar columns. Data-dependent acquisition (DDA), wide-window acquisition (WWA), data-independent acquisition (DIA), and commonly used advanced data analysis algorithms were benchmarked. Using DDA, 1790 proteins covering a dynamic range of four orders of magnitude were identified in a single cell. Using DIA, proteome coverage increased to more than 2200 proteins identified from single-cell level input in a 20 min active gradient. The workflow enabled differentiation of two cell lines, demonstrating its suitability to cellular heterogeneity determination.

In Vivo Electrochemical Biosensors: Recent Advances in Molecular Design, Electrode Materials, and Electrochemical Devices.

Abstract: Electrochemical biosensors provide powerful tools for dissecting the dynamically changing neurochemical signals in the living brain, which contribute to the insight into the physiological and pathological processes of the brain, due to their high spatial and temporal resolutions. Recent advances in the integration of in vivo electrochemical sensors with cross-disciplinary advances have reinvigorated the development of in vivo sensors with even better performance. In this Review, we summarize the recent advances in molecular design, electrode materials, and electrochemical devices for in vivo electrochemical sensors from molecular to macroscopic dimensions, highlighting the methods to obtain high performance for fulfilling the requirements for determination in the complex brain through flexible and smart design of molecules, materials, and devices. Also, we look forward to the development of next-generation in vivo electrochemical biosensors.

MALDI-IHC-Guided In-Depth Spatial Proteomics: Targeted and Untargeted MSI Combined

Abstract: Recently, a novel technology was published, utilizing the strengths of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) and immunohistochemistry (IHC), achieving highly multiplexed, targeted imaging of biomolecules in tissue. This new technique, called MALDI-IHC, opened up workflows to target molecules of interest using MALDI-MSI that are usually targeted by standard IHC. In this paper, the utility of targeted MALDI-IHC and its complementarity with untargeted on-tissue bottom-up spatial proteomics is explored using breast cancer tissue. Furthermore, the MALDI-2 effect was investigated and demonstrated to improve MALDI-IHC. Formalin-fixed paraffin-embedded (FFPE) human breast cancer tissue sections were stained for multiplex MALDI-IHC with six photocleavable mass-tagged (PC-MT) antibodies constituting a breast cancer antibody panel (CD20, actin-αSM, HER2, CD68, vimentin, and panCK). K-means spatial clusters were created based on the MALDI-IHC images and cut out using laser-capture microdissection (LMD) for further untargeted LC-MS-based bottom-up proteomics analyses. Numerous peptides could be tentatively assigned to multiple proteins, of which three proteins were also part of the antibody panel (vimentin, keratins, and actin). Post-ionization with MALDI-2 showed an increased intensity of the PC-MTs and suggests options for the development of new mass-tags. Although the on-tissue digestion covered a wider range of proteins, the MALDI-IHC allowed for easy and straightforward identification of proteins that were not detected in untargeted approaches. The combination of the multiplexed MALDI-IHC with image-guided proteomics showed great potential to further investigate diseases by providing complementary information from the same tissue section and without the need for customized instrumentation.

Rapid Approximate Subset-Based Spectra Prediction for Electron Ionization–Mass Spectrometry

Abstract: Mass spectrometry is a vital tool in the analytical chemist’s toolkit, commonly used to identify the presence of known compounds and elucidate unknown chemical structures. All of these applications rely on having previously measured spectra for known substances. Computational methods for predicting mass spectra from chemical structures can be used to augment existing spectral databases with predicted spectra from previously unmeasured molecules. In this paper, we present a method for prediction of electron ionization–mass spectra (EI–MS) of small molecules that combines physically plausible substructure enumeration and deep learning, which we term rapid approximate subset-based spectra prediction (RASSP). The first of our two models, FormulaNet, produces a probability distribution over chemical subformulae to achieve a state-of-the-art forward prediction accuracy of 92.9% weighted (Stein) dot product and database lookup recall (within top 10 ranked spectra) of 98.0% when evaluated against the NIST 2017 Mass Spectral Library. The second model, SubsetNet, produces a probability distribution over vertex subsets of the original molecule graph to achieve similar forward prediction accuracy and superior generalization in the high-resolution, low-data regime. Spectra predicted by our best model improve upon the previous state-of-the-art spectral database lookup error rate by a factor of 2.9×, reducing the lookup error (top 10) from 5.7 to 2.0%. Both models can train on and predict spectral data at arbitrary resolution. Source code and predicted EI–MS spectra for 73.2M small molecules from PubChem will be made freely accessible online.

Direct and Specific Detection of Glyphosate Using a Phosphatase-like Nanozyme-Mediated Chemiluminescence Strategy.

Abstract: Most organophosphorus pesticide (OP) sensors reported in the literature rely on the inhibition effect of OPs on the activity of acetylcholinesterase (AChE), which suffer from the drawbacks of lack of selective recognition of OPs, high cost, and poor stability. Herein, we proposed a novel chemiluminescence (CL) strategy for the direct detection of glyphosate (an organophosphorus herbicide) with high sensitivity and specificity, which is based on the porous hydroxy zirconium oxide nanozyme (ZrOX-OH) obtained via a facile alkali solution treatment of UIO-66. ZrOX-OH displayed excellent phosphatase-like activity, which could catalyze the dephosphorylation of 3-(2'-spiroadamantyl)-4-methoxy-4-(3'-phosphoryloxyphenyl)-1,2-dioxetane (AMPPD) to generate strong CL. The experimental results showed that the phosphatase-like activity of ZrOX-OH is closely related to the content of hydroxyl groups on their surface. Interestingly, ZrOX-OH with phosphatase-like properties exhibited a unique response to glyphosate because of the consumption of the surface hydroxyl group by the unique carboxyl group of glyphosates and was thus employed to develop a CL sensor for direct and selective detection of glyphosate without using bio-enzymes. The recovery for glyphosate detection of cabbage juice ranged from 96.8 to 103.0%. We believe that the as-proposed CL sensor based on ZrOX-OH with phosphatase-like properties supplies a simpler and more highly selective approach for OP assay and provides a new method for the development of CL sensors for the direct analysis of OPs in real samples.

Emerging Wearable Chemical Sensors Enabling Advanced Integrated Systems toward Personalized and Preventive Medicine.

Abstract: ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTEmerging Wearable Chemical Sensors Enabling Advanced Integrated Systems toward Personalized and Preventive MedicineTianyiyi HeTianyiyi HeDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, SingaporeMore by Tianyiyi HeView Biography, Feng WenFeng WenDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, SingaporeMore by Feng WenView Biography, Yanqin YangYanqin YangDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, SingaporeMore by Yanqin YangView Biography, Xianhao LeXianhao LeDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, SingaporeMore by Xianhao LeView Biography, Weixin LiuWeixin LiuDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, SingaporeMore by Weixin LiuView Biographyhttps://orcid.org/0000-0002-5526-3487, and Chengkuo Lee*Chengkuo LeeDepartment of Electrical and Computer Engineering, National University of Singapore, 4 Engineering Drive 3, Singapore 117583, SingaporeCenter for Intelligent Sensors and MEMS, National University of Singapore, Block E6 #05-11, 5 Engineering Drive 1, Singapore 117608, Singapore*[email protected]More by Chengkuo LeeView Biographyhttps://orcid.org/0000-0002-8886-3649Cite this: Anal. Chem. 2023, 95, 1, 490–514Publication Date (Web):January 10, 2023Publication History Received14 October 2022Published online10 January 2023Published inissue 10 January 2023https://doi.org/10.1021/acs.analchem.2c04527Copyright © 2023 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views419Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (5 MB) Get e-AlertsSUBJECTS:Anatomy,Carbohydrates,Sensors,Substrates,Wearable electronics Get e-Alerts

Homogeneous Electrochemical Immunoassay Using an Aggregation-Collision Strategy for Alpha-Fetoprotein Detection.

Abstract: Homogeneous immunoassays represent an attractive alternative to traditional heterogeneous assays due to their simplicity and high efficiency. Homogeneous electrochemical assays, however, are not commonly accessed due to the requirement of electrode immobilization of the recognition elements. Herein, we demonstrate a new homogeneous electrochemical immunoassay based on the aggregation-collision strategy for the quantification of tumor protein biomarker alpha-fetoprotein (AFP). The detection principle relies on the aggregation of AgNPs induced by the molecular biorecognition between AFP and AgNPs-anti-AFP probes, which leads to an increased AgNP size and decreased AgNP concentration, allowing an accurate self-validated dual-mode immunoassay by performing nanoimpact electrochemistry (NIE) of the oxidation of AgNPs. The intrinsic one-by-one analytical capability of NIE as well as the participation of all of the atoms of the AgNPs in signal transduction greatly elevates the detection sensitivity. Accordingly, the current sensor enables a limit of detection (LOD) of 5 pg/mL for AFP analysis with high specificity and efficiency. More importantly, reliable detection of AFP in diluted human sera of hepatocellular carcinoma (HCC) patients is successfully achieved, indicating that the NIE-based homogeneous immunoassay shows great potential in HCC liquid biopsy.

Finding the Lost Dissociation Constant of Electrochemical Aptamer-Based Biosensors.

Abstract: Electrochemical aptamer-based (E-AB) biosensors afford real-time measurements of the concentrations of molecules directly in complex matrices and in the body, offering alternative strategies to develop innovative personalized medicine tools. While different electroanalytical techniques have been used to interrogate E-AB sensors (i.e., cyclic voltammetry, electrochemical impedance spectroscopy, and chronoamperometry) to resolve the change in electron transfer of the aptamer's covalently attached redox reporter, square-wave voltammetry remains a widely used technique due to its ability to maximize the redox reporter's faradic contribution to the measured current. Several E-AB sensors interrogated with this technique, however, show lower aptamer affinity (i.e., μM-mM) even in the face of employing aptamers that have high affinities (i.e., nM-μM) when characterized using solution techniques such as isothermal titration calorimetry (ITC) or fluorescence spectroscopy. Given past reports showing that E-AB sensor's response is dependent on square-wave interrogation parameters (i.e., frequency and amplitude), we hypothesized that the difference in dissociation constants measured with solution techniques stemmed from the electrochemical interrogation technique itself. In response, we decided to compare six dissociation constants of aptamers when characterized in solution with ITC and when interrogated on electrodes with electrochemical impedance spectroscopy, a technique able to, in contrast to square-wave voltammetry, deconvolute and quantify E-AB sensors' contributions to the measured current. In doing so, we found that we were able to measure dissociation constants that were either separated by 2-3-fold or within experimental errors. These results are in contrast with square-wave voltammetry-measured dissociation constants that are at the most separated by 2-3 orders of magnitude from ones measured by ITC. We thus envision that the versatility and time scales covered by electrochemical impedance spectroscopy offer the highest sensitivity to measure target binding in electrochemical biosensors relying on changes in electron-transfer rates.

Upregulation of Formaldehyde in Parkinson's Disease Found by a Near-Infrared Lysosome-Targeted Fluorescent Probe.

Abstract: Parkinson's disease (PD) is one of the major neurodegenerative diseases caused by complex pathological processes. As a signal molecule, formaldehyde is closely linked to nervous systems, but the relationship between PD and formaldehyde levels remains largely unclear. We speculated that formaldehyde might be a potential biomarker for PD. To prove it, we constructed the first near-infrared (NIR) lysosome-targeted formaldehyde fluorescent probe (named NIR-Lyso-FA) to explore the relationship between formaldehyde and PD. The novel fluorescent probe achieves formaldehyde detection in vitro and in vivo, thanks to its excellent properties such as NIR emission, large Stokes shift, and fast response to formaldehyde. Crucially, utilizing the novel probe NIR-Lyso-FA, formaldehyde overexpression was discovered for the first time in cellular, zebrafish, and mouse PD models, supporting our guess that formaldehyde can function as a possible biomarker for PD. We anticipate that this finding will offer insightful information for PD pathophysiology, diagnosis, medication development, and treatment.

One Stone, Three Birds: A Smart Single Fluorescent Probe for Simultaneous and Discriminative Imaging of Lysosomes, Lipid Droplets, and Mitochondria.

Abstract: Complex intracellular life processes are usually completed through the cooperation of multiple organelles. Real-time tracking of the interplays between multiple organelles with a single fluorescent probe (SFP) is very helpful to deepen our understanding of complex biological processes. So far, SFP for simultaneously differentiating and visualizing of more than two different organelles has not been reported. Herein, we report an SFP (named ICM) that can be used for simultaneously differentiating and visualizing three important organelles: mitochondria, lysosomes, and lipid droplets (LDs). The probe can simultaneously light up mitochondria/lysosomes (∼700 nm) and LDs (∼480 nm) at significantly different emission wavelengths with high fidelity, and mitochondria and lysosomes can be effectively distinguished by their different shapes and fluorescence intensities. With this smart probe, real-time and simultaneous tracking of the interplays of these three organelles was successfully achieved for the first time.

Dual-Gene-Controlled Rolling Circle Amplification Strategy for SARS-CoV-2 Analysis

Abstract: The development of sensitive, accurate, and conveniently operated methods for the simultaneous assay of two nucleic acids is promising while still challenging. In this work, by using two genes (the N gene and RdRp gene) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as examples, we have designed an ingenious dual-gene-controlled rolling circle amplification (RCA) strategy to propose an accurate and sensitive electrochemical method. Specifically, the coexistence of the two target genes can trigger the RCA reaction to generate a number of repeated G-quadruplex (G4)-forming sequences. These sequences then switch into G4/hemin complexes with redox activity after the incubation of hemin, which can catalyze the TMB/H2O2 substrates to produce significantly enhanced current responses. Experimental results reveal that the proposed method exhibits satisfying feasibility and analytical performance, enabling the sensitive detection of SARS-CoV-2 in the range of 0.1–5000 pM, with the detection limit of 57 fM. Meanwhile, because only the simultaneous existence of the two target genes can effectively trigger the downstream amplification reaction, this method can effectively avoid false-positives and ensure specificity as well as accuracy. Furthermore, our method can distinguish the COVID-19 samples from healthy people, and the outcomes show a satisfying agreement with the results of RT-PCR, manifesting that our label-free dual-gene-controlled RCA strategy exhibits great possibility in clinical application.

Progression of Paper-Based Point-of-Care Testing toward Being an Indispensable Diagnostic Tool in Future Healthcare.

Abstract: Point-of-care (POC) diagnostics in particular focuses on the timely identification of harmful conditions close to the patients' needs. For future healthcare these diagnostics could be an invaluable tool especially in a digitalized or telemedicine-based system. However, while paper-based POC tests, with the most prominent example being the lateral flow assay (LFA), have been especially successful due to their simplicity and timely response, the COVID-19 pandemic highlighted their limitations, such as low sensitivity and ambiguous responses. This perspective discusses strategies that are currently being pursued to evolve such paper-based POC tests toward a superior diagnostic tool that provides high sensitivities, objective result interpretation, and multiplexing options. Here, we pinpoint the challenges with respect to (i) measurability and (ii) public applicability, exemplified with select cases. Furthermore, we highlight promising endeavors focused on (iii) increasing the sensitivity, (iv) multiplexing capability, and (v) objective evaluation to also ready the technology for integration with machine learning into digital diagnostics and telemedicine. The status quo in academic research and industry is outlined, and the likely highly relevant role of paper-based POC tests in future healthcare is suggested.

Ion-Exchange Chromatography Coupled to Mass Spectrometry in Life Science, Environmental, and Medical Research

Abstract: anion-exchange chromatography. The column’s stationary phase is positively charged and equilibrated with a mobile phase containing negative ions (e.g., OH − or another anion) at a minimal concentration. When a sample is introduced with negatively charged or polarizable analytes, the charged analytes displace the negative ions (e.g., OH − ions) from the stationary phase and bind instead, therefore being retained. Analyte ions have differential affinity for the stationary phase depending upon their charge; the affinity is directly determined by Coulombic force. The ionic strength of the mobile phase is usually increased for a gradient elution where the concentration of ions in the mobile phase (e.g., OH − ) is gradually increased until the analyte ions are displaced by the increasing ion concentration in the mobile phase (isocratic elution is also sometimes used)

On the Road from Single-Atom Materials to Highly Sensitive Electrochemical Sensing and Biosensing.

Abstract: ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTOn the Road from Single-Atom Materials to Highly Sensitive Electrochemical Sensing and BiosensingLei JiaoLei JiaoKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. ChinaInstitute of Molecular Metrology, College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071, P. R. ChinaMore by Lei JiaoView Biography, Weiqing XuWeiqing XuKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. ChinaMore by Weiqing XuView Biography, Yu WuYu WuKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. ChinaMore by Yu WuView Biography, Hengjia WangHengjia WangKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. ChinaMore by Hengjia WangView Biography, Liuyong HuLiuyong HuHubei Engineering Technology Research Center of Optoelectronic and New Energy Materials, Hubei Key Laboratory of Plasma Chemistry and Advanced Materials, School of Materials Science and Engineering, Wuhan Institute of Technology, Wuhan 430205, P. R. ChinaMore by Liuyong HuView Biography, Wenling GuWenling GuKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. ChinaMore by Wenling GuView Biography, and Chengzhou Zhu*Chengzhou ZhuKey Laboratory of Pesticide and Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensing Technology and Health, College of Chemistry, Central China Normal University, Wuhan, Hubei 430079, P. R. China*[email protected]More by Chengzhou ZhuView Biographyhttps://orcid.org/0000-0003-0679-7965Cite this: Anal. Chem. 2023, 95, 1, 433–443Publication Date (Web):January 10, 2023Publication History Received19 April 2022Published online10 January 2023Published inissue 10 January 2023https://doi.org/10.1021/acs.analchem.2c01722Copyright © 2023 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views1178Altmetric-Citations2LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (5 MB) Get e-AlertscloseSUBJECTS:Biotechnology,Electrochemical detection,Metals,Platinum,Sensors Get e-Alerts

Chromatographic Phospholipid Trapping for Automated H/D Exchange Mass Spectrometry of Membrane Protein–Lipid Assemblies

Abstract: Lipid interactions modulate the function, folding, structure, and organization of membrane proteins. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) has emerged as a useful tool to understand the structural dynamics of these proteins within lipid environments. Lipids, however, have proven problematic for HDX-MS analysis of membrane-embedded proteins due to their presence of impairing proteolytic digestion, causing liquid chromatography column fouling, ion suppression, and/or mass spectral overlap. Herein, we describe the integration of a chromatographic phospholipid trap column into the HDX-MS apparatus to enable online sample delipidation prior to protease digestion of deuterium-labeled protein–lipid assemblies. We demonstrate the utility of this method on membrane scaffold protein–lipid nanodisc—both empty and loaded with the ∼115 kDa transmembrane protein AcrB—proving efficient and automated phospholipid capture with minimal D-to-H back-exchange, peptide carry-over, and protein loss. Our results provide insights into the efficiency of phospholipid capture by ZrO2-coated and TiO2 beads and describe how solution conditions can be optimized to maximize not only the performance of our online but also the existing offline, delipidation workflows for HDX-MS. We envision that this HDX-MS method will significantly ease membrane protein analysis, allowing to better interrogate their dynamics in artificial lipid bilayers or even native cell membranes.

Amorphous Fe-Containing Phosphotungstates Featuring Efficient Peroxidase-like Activity at Neutral pH: Toward Portable Swabs for Pesticide Detection with Tandem Catalytic Amplification.

Abstract: Peroxidase-mimetic materials are intensively applied to establish multienzyme systems because of their attractive merits. However, almost all of the nanozymes explored exhibit catalytic capacity only under acidic conditions. The pH mismatch between peroxidase mimics in acidic environments and bioenzymes under neutral conditions significantly restricts the development of enzyme-nanozyme catalytic systems especially for biochemical sensing. To solve this problem, here amorphous Fe-containing phosphotungstates (Fe-PTs) featuring high peroxidase activity at neutral pH were explored to fabricate portable multienzyme biosensors for pesticide detection. The strong attraction of negatively charged Fe-PTs to positively charged substrates as well as the accelerated regeneration of Fe2+ by the Fe/W bimetallic redox couples was demonstrated to play important roles in endowing the material with peroxidase-like activity in physiological environments. Consequently, integrating the developed Fe-PTs with acetylcholinesterase and choline oxidase led to an enzyme-nanozyme tandem platform with good catalytic efficiency at neutral pH for organophosphorus pesticide response. Furthermore, they were immobilized onto common medical swabs to fabricate portable sensors for paraoxon detection conveniently based on smartphone sensing, showing excellent sensitivity, good anti-interference capacity, and low detection limit (0.28 ng/mL). Our contribution expands the horizon of acquiring peroxidase activity at neutral pH, and it will also open avenues to construct portable and effective biosensors for pesticides and other analytes.

Enhanced Luminol Chemiluminescence with Oxidase-like Properties of FeOOH Nanorods for the Sensitive Detection of Uric Acid.

Abstract: FeOOH nanorods, as one-dimensional nanomaterials, have been widely used in many fields due to their stable properties, low cost, and easy synthesis, but their application in the field of chemiluminescence (CL) is rarely reported. In this work, FeOOH nanorods were synthesized by a simple and environmentally friendly one-pot hydrothermal method and used for the first time as a catalyst for generating strong CL with luminol without additional oxidant. Remarkably, luminol-FeOOH exhibits about 250 times stronger CL than the luminol-H2O2 system. Its CL intensity was significantly quenched by uric acid. We established a simple, rapid, sensitive, and selective CL method for the detection of uric acid with a linear range of 20-1000 nM and a detection limit of 6.3 nM (S/N = 3). In addition, we successfully applied this method to the detection of uric acid in human serum, and the standard recoveries were 95.6-106.4%.

Ultrasensitive CRISPR/Cas13a-Mediated Photoelectrochemical Biosensors for Specific and Direct Assay of miRNA-21.

Abstract: Sensitive and specific assay of microRNAs (miRNAs) is beneficial to early disease screening. Herein, we for the first time proposed clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a-mediated photoelectrochemical biosensors for the direct assay of miRNA-21. In this study, compared with traditional nucleic acid-based signal amplification strategies, the CRISPR/Cas13a system can greatly improve the specificity and sensitivity of target determination due to its accurate recognition and high-efficient trans-cleavage capability without complex nucleic acid sequence design. Moreover, compared with the CRISPR/Cas12a-based biosensing platform, the developed CRISPR/Cas13a-mediated biosensor can directly detect RNA targets without signal transduction from RNA to DNA, thereby avoiding signal leakage and distortion. Generally, the proposed biosensor reveals excellent analysis capability with a wider linear range from 1 fM to 5 nM and a lower detection limit of 1 fM. Additionally, it also shows satisfactory stability in the detection of human serum samples and cell lysates, manifesting that it has great application prospects in the areas of early disease diagnosis and biomedical research.

Schiff-Base Chemistry-Coupled Catechol Oxidase-Like Nanozyme Reaction as a Universal Sensing Mode for Ultrasensitive Biosensing.

Abstract: Expanding sensing modes and improving catalytic performance of nanozyme-based analytical chemistry are beneficial to realizing the desired biosensing of analytes. Herein, Schiff-base chemistry coupled with a novel catechol oxidase-like nanozyme (CHzyme) is designed and constructed, exhibiting two main advantages, including (1) improving catalytic performance by nearly 2-fold compared with only the oxidase-like role of CHzyme; (2) increasing the designability of the output signal by signal transduction of cascade reaction. Thereafter, the substrate sensing modes based on a cascade reaction between the CHzyme-catalyzed reaction and Schiff-base chemistry are proposed and comprehensively studied, containing catalytic substrate sensing mode, competitive substrate sensing mode, and generated substrate sensing mode, expecting to be employed in environmental monitoring, food analyses, and clinical diagnoses, respectively. More meaningfully, the generated substrate sensing mode is successfully applied to construct a cascade reaction coupling ratiometric fluorescent immunoassay for the detection of clenbuterol, increasing 15-fold in detection sensitivity compared with the traditional enzyme-linked immunosorbent assay. It is expected that the expanded universal substrate sensing modes and the Schiff-base chemistry-enhanced nanozyme can enlighten the exploration of innovative biosensors.

Microfluidic Separation and Enrichment of Escherichia coli by Size Using Viscoelastic Flows.

Abstract: Here, we achieve the separation and enrichment of Escherichia coli clusters from its singlets in a viscoelastic microfluidic device. E. coli, an important prokaryotic model organism and a widely used microbial factory, can aggregate in clusters, leading to biofilm development that can be detrimental to human health and industrial processes. The ability to obtain high-purity populations of E. coli clusters is of significance for biological, biomedical, and industrial applications. In this study, polystyrene particles of two different sizes, 1 and 4.8 μm, are used to mimic E. coli singlets and clusters, respectively. Experimental results show that particles migrate toward the channel center in a size-dependent manner, due to the combined effects of inertial and elastic forces; 4.8 and 1 μm particles are found to have lateral equilibrium positions closer to the channel centerline and sidewalls, respectively. The size-dependent separation performance of the microdevice is demonstrated to be affected by three main factors: channel length, the ratio of sheath to sample flow rate, and poly(ethylene oxide) (PEO) concentration. Further, the separation of E. coli singlets and clusters is achieved at the outlets, and the separation efficiency is evaluated in terms of purity and enrichment factor.

Ratiometric Detection of Ochratoxin A Using a Regenerable COF-Au-MB-Apt Signal Probe on a Thermal-Regulated Sensor Module.

Abstract: Ochratoxin A (OTA) frequently contaminates grains and consequently threatens human health. Herein, we develop a regenerable signal probe and apply it to a Au-based screen-printed electrode module (SPE) for OTA determination. The signal probe, containing a structural covalent organic framework, gold nanoparticles (AuNPs), indicative methylene blue (MB), and a highly selective aptamer, is synthesized with hydrothermal and self-assembly methods. The SPE is permanently functionalized with Prussian blue (PB), AuNPs, and semicomplementary ssDNA. The signal probe, absorbed onto this SPE via hybridization, is competitively expelled by OTA, providing a ratiometric readout of ΔIMB/IPB. Probe regeneration, to erase expired COF-Au-MB-Apt after each analysis, is established with the synergy of OTA-conducted Apt-ssDNA dissociation and on-chip thermal regulation. This advantage powerfully guarantees reduplicative analyses by avoiding irreversible Apt-OTA combination and accumulation on the sensing interface. Regenerations are performed in repetitive cycles (N = 7) with 98.5% reproduction efficiency, and IMB and IPB fluctuations are calculated as 1.45 and 1.12%. This method shows log-linear OTA response in a wide range from 0.2 pg/mL to 0.6 μg/mL, and the limit of detection is 0.12 pg/mL. During natural OTA determinations, recommended readouts match well with HPLC with less than 4.82% relative error.

Fluorescence Imaging of Intracellular Glutathione Levels in the Endoplasmic Reticulum to Reveal the Inhibition Effect of Rutin on Ferroptosis.

Abstract: Ferroptosis is an emerging form of nonapoptotic cell death, and the search for novel ferroptosis inhibitors is of great importance to explore unique cytoprotective strategies against ferroptosis-relevant diseases. In this work, we present an endoplasmic reticulum-targeting fluorescent probe (ER-G) for the imaging of intracellular glutathione (GSH) levels and revealed the inhibition effect of rutin on ferroptosis. Structurally, ER-G utilized a cyclohexyl sulfonylurea as the endoplasmic reticulum-targeting unit, and single-crystal X-ray diffraction analysis confirmed that ER-G possessed a N-oxide pyridine sulfinyl group instead of sulfone. After the response of ER-G to GSH, the fluorescence intensity at 523 nm displayed a significant increase by 3900-fold. ER-G showed extreme sensitivity and selectivity to GSH. The fluorescence imaging results demonstrated that ER-G exhibited excellent endoplasmic reticulum-targeting properties and could be applied to monitor GSH levels in the endoplasmic reticulum during the erastin-induced ferroptosis process. By the fluorescence imaging of GSH levels in the endoplasmic reticulum, it was demonstrated that rutin could efficiently block the depletion of GSH during erastin-induced ferroptosis and potentially act as a novel ferroptosis inhibitor. Moreover, unlike traditional ferroptosis inhibitors, it was speculated that the inhibition mechanism of rutin to ferroptosis was the integration of the chelate effect on Fe(II) ions and antioxidant effect. We expect that fluorescence imaging of GSH levels in the endoplasmic reticulum could provide a convenient and feasible method to evaluate the inhibition effect of small molecules on ferroptosis.

NMR Metabolomics Methods for Investigating Disease.

Abstract: ADVERTISEMENT RETURN TO ISSUEPREVReviewNEXTNMR Metabolomics Methods for Investigating DiseaseG. A. Nagana GowdaG. A. Nagana GowdaNorthwest Metabolomics Research Center and Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, Washington 98109, United StatesMore by G. A. Nagana GowdaView Biographyhttps://orcid.org/0000-0002-0544-7464 and Daniel Raftery*Daniel RafteryNorthwest Metabolomics Research Center and Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, Washington 98109, United StatesFred Hutchinson Cancer Research Center, Seattle, Washington 98109, United States*Tel: 206-543-9709. Fax: 206-616-4819. Email: [email protected]More by Daniel RafteryView Biographyhttps://orcid.org/0000-0003-2467-8118Cite this: Anal. Chem. 2023, 95, 1, 83–99Publication Date (Web):January 10, 2023Publication History Received18 October 2022Published online10 January 2023Published inissue 10 January 2023https://doi.org/10.1021/acs.analchem.2c04606Copyright © 2023 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views329Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (8 MB) Get e-AlertsSUBJECTS:Anatomy,COVID-19,Metabolism,Metabolomics,Nuclear magnetic resonance spectroscopy Get e-Alerts

In Situ and Quantitatively Monitoring the Dynamic Process of Ferroptosis in Single Cancer Cells by Scanning Electrochemical Microscopy.

Abstract: Ferroptosis, as a promising therapeutic strategy for cancers, has aroused great interest. Quantifying the quick dynamic changes in key parameters during the early course of ferroptosis can provide insights for understanding the underlying mechanisms of ferroptosis and help the development of therapies targeting ferroptosis. However, in situ and quantitatively monitoring the quick responses of living cancer cells to ferroptosis at the single-cell level remains technically challenging. In this work, we selected HuH7 cells (hepatocellular carcinoma (HCC) cells) as a cell model and Erastin as a typical ferroptosis inducer. We utilized scanning electrochemical microscopy (SECM) to quantitatively and in situ monitor the early course of ferroptosis in HuH7 cells by characterizing the three key parameters of cell ferroptosis (i.e., cell membrane permeability, respiratory activity, and the redox state). The SECM results show that the membrane permeability of ferroptotic HuH7 cells continuously increased from 0 to 8.1 × 10-5 m s-1, the cellular oxygen consumption was continuously reduced by half, and H2O2 released from the cells exhibited periodic bursts during the early course of ferroptosis, indicating the gradually destroyed cell membrane structure and intensified oxidative stress. Our work realizes, for the first time, the in situ and quantitative monitoring of the cell membrane permeability, respiratory activity, and H2O2 level of the early ferroptosis process of a single living cancer cell with SECM, which can contribute to the understanding of the physiological process and underlying mechanisms of ferroptosis.

Explaining the Decay of Nucleic Acid-Based Sensors under Continuous Voltammetric Interrogation.

Abstract: Nucleic acid-based electrochemical sensors (NBEs) can support continuous and highly selective molecular monitoring in biological fluids, both in vitro and in vivo, via affinity-based interactions. Such interactions afford a sensing versatility that is not supported by strategies that depend on target-specific reactivity. Thus, NBEs have significantly expanded the scope of molecules that can be monitored continuously in biological systems. However, the technology is limited by the lability of the thiol-based monolayers employed for sensor fabrication. Seeking to understand the main drivers of monolayer degradation, we studied four possible mechanisms of NBE decay: (i) passive desorption of monolayer elements in undisturbed sensors, (ii) voltage-induced desorption under continuous voltammetric interrogation, (iii) competitive displacement by thiolated molecules naturally present in biofluids like serum, and (iv) protein binding. Our results indicate that voltage-induced desorption of monolayer elements is the main mechanism by which NBEs decay in phosphate-buffered saline. This degradation can be overcome by using a voltage window contained between -0.2 and 0.2 V vs Ag|AgCl, reported for the first time in this work, where electrochemical oxygen reduction and surface gold oxidation cannot occur. This result underscores the need for chemically stable redox reporters with more positive reduction potentials than the benchmark methylene blue and the ability to cycle thousands of times between redox states to support continuous sensing for long periods. Additionally, in biofluids, the rate of sensor decay is further accelerated by the presence of thiolated small molecules like cysteine and glutathione, which can competitively displace monolayer elements even in the absence of voltage-induced damage. We hope that this work will serve as a framework to inspire future development of novel sensor interfaces aiming to eliminate the mechanisms of signal decay in NBEs.

Dry Chemistry-Based Bipolar Electrochemiluminescence Immunoassay Device for Point-of-Care Testing of Alzheimer-Associated Neuronal Thread Protein.

Abstract: In this study, we developed, for the first time, a novel dry chemistry-based bipolar electrochemiluminescence (ECL) immunoassay device for point-of-care testing (POCT) of Alzheimer-associated neuronal thread protein (AD7c-NTP), where the ECL signals were automatically collected and analyzed after the sample and buffer solutions were manually added onto the immunosensor. The proposed immunoassay device contains an automatic ECL analyzer and a dry chemistry-based ECL immunosensor fabricated with a screen-printed fiber material-based chip and a three-dimensional (3D)-printed shell. Each pad of the fiber material-based chip was premodified with certain reagents for immunoreaction and then assembled to form the ECL immunosensor. The self-enhanced ECL of the Ru(II)-poly-l-lysine complex and the lateral flow fiber material-based chip make the addition of coreactants and repeated flushing unnecessary. Only the sample and buffer solutions are added to the ECL immunosensor, and the process of ECL detection can be completed in about 6 min using the proposed automatic ECL analyzer. Under optimized conditions, the linear detection range for AD7c-NTP was 1 to 104 pg/mL, and the detection limit was 0.15 pg/mL. The proposed ECL immunoassay device had acceptable selectivity, stability, and reproducibility and had been successfully applied to detect AD7c-NTP levels in human urine. In addition, the accurate detection of AD7c-NTP and duplex detection of AD7c-NTP and apolipoprotein E ε4 gene were also validated. It is believed that the proposed ECL immunoassay device may be a candidate for future POCT applications.